Anti-Mannose Receptor antibody (ab64693)

IHC image of ab64693 staining in human lung formalin fixed paraffin embedded tissue section*, performed on a Leica Bond^TM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab64693, 0.1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

Paraffin embedded, formalin-fixed mouse lung tissue labelling the mannose receptor with ab64693 at 1/500 in immunohistochemical analysis. An Alexa Fluor^® 647 Goat anti-rabbit IgG was used as the secondary antibody, this was artificially colored green by software to enhance stain visibility. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. 15% serum was used as blocking agent for 1 hour at room temperature.

ab64693 staining Mannose Receptor in MOLT4 cells. The cells were fixed with 100% methanol (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions.  The cells were then incubated with the antibody ab64693 at 1µg/ml and ab7291 (Mouse monoclonal to alpha Tubulin - Loading Control) used at a 1/1000 dilution overnight at +4°C. The secondary antibodies were ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed, (pseudo-colored green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594) preadsorbed, (colored red), both used at a 1/1000 dilution for 1 hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43 µM for 1hour at room temperature.

ab64693 staining Mannose Receptor in Mouse heart tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with acetone and blocked with 0.075% H₂O₂ for 10 minutes at room temperature. Samples were incubated with primary antibody (1/100 in 1x PBS + 10% heat inactivated FCS) for 1 hour. An undiluted HRP-conjugated Rabbit anti-rabbit IgG polyclonal was used as the secondary antibody.

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